Macrophages polarize to the pro-inflammatory phenotype and delay neutrophil efferocytosis to augment Aggregatibacter actinomycetemcomitance JP2 clearance.

OBJECTIVE: The study examines how neutrophils cross-talk with macrophages during JP2 Aggregatibacter actinomycetemcomitance infection and factors that are involved in inflammatory resolution and efferocytosis. BACKGROUND: Although sub-gingival bacteria constitute the primary initiating factor in the pathogenesis of molar-incisor pattern periodontitis (MIPP), the non-resolved host response has a major role in tissue destruction. While evidence links neutrophils to MIPP pathogenesis, their clearance during inflammatory resolution, governed by macrophages, is poorly understood. METHODS: Human neutrophils (differentiated from HL60 cells) and macrophages (differentiated from THP1 cells) were inoculated with JP2. The supernatants were collected and exposed to naïve neutrophils or macrophages with or without exposure to JP2. Reactive oxygen species (ROS) were measured with 2'-7'-dichlorofluorescein-diacetate and a fluorescent plate reader. Immunofluorescence labeling of CD47 and cell vitality were examined using flow cytometry. Macrophage polarization was tested by immunofluorescence staining for CD163 and CD68 and a fluorescent microscope, and TNFα and IL-10 secretion was tested using ELISA and RT-PCR. Efferocytosis was examined by pHrodo and carboxyfluorescein succinimidyl ester staining and fluorescent microscopy. In vivo, macrophages were depleted from C57Bl/6 mice and neutrophil CD47 levels were tested using the subcutaneous chamber model. RESULTS: Neutrophils exposed to macrophage supernatant show increased ROS, mainly extracellularly, that increased during JP2 infection. Macrophages showed pro-inflammatory M1 phenotype polarization during JP2 infection, and their supernatants prolonged neutrophil survival by inhibiting CD47 down-expression and reducing neutrophil necrosis and apoptosis. Also, the macrophages delay neutrophil efferocytosis during JP2 infection which, in turn, enhanced JP2 clearance. Depletion of macrophages in mice mildly prevented neutrophils CD47 reduction and reduced JP2 clearance. The JP2 infection in mice also led to macrophage M1 polarization similar to the in vitro results. CONCLUSIONS: As shown in this study, neutrophil efferocytosis potentially may be reduced during JP2 infection, promoting JP2 clearance, which may contribute to the inflammatory-mediated periodontal tissue damage.

Mechanical force application and inflammation induce osteoclastogenesis by independent pathways.

OBJECTIVE: To investigate the functional changes of PDL fibroblasts in the presence of mechanical force, inflammation, or a combination of force and inflammation. MATERIALS AND METHODS: Inflammatory supernatants were prepared by inoculating human neutrophils with Porphyromonas gingivalis. Primary human PDL fibroblasts (PDLF), gingival fibroblasts (GFs), and osteoblasts (Saos2) were then exposed to the inflammatory supernatants. Orthodontic force on the PDLFs was simulated by centrifugation. Analyses included cell proliferation, cell viability, cell cycle, and collagen expression, as well as osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-Β ligand (RANKL) expression. RESULTS: Mechanical force did not affect PDLF viability, but it increased the metabolic rate compared to resting cells. Force application shifted the PDLF cell cycle to the G0/G1 phase, arresting cell proliferation and leading to elevated collagen production, mild OPG level elevation, and robust RANKL level elevation. Including an inflammatory supernatant in the presence of force did not affect PDLF viability, proliferation, or cytokine expression. By contrast, the inflammatory supernatant increased RANKL expression in GFs, but not in Saos2 cells. CONCLUSION: Applying mechanical force significantly affects PDLF function. Although inflammation had no effect on PDLF or Saos2 cells, it promoted RANKL expression in GF cells. Within the limitations of the in vitro model, the results suggest that periodontal inflammation and mechanical forces could affect bone catabolism through effects on different cell types, which may culminate in synergistic bone resorption.

Enterococcus faecalis sustained infection induces macrophage pro-resolution polarization.

AIM: To investigate macrophage function in the presence of sustained infection with Enterococcus faecalis, a prevalent root canal resident in asymptomatic apical periodontitis. METHODOLOGY: The human monocyte cell line (THP-1) was differentiated into macrophages by exposure to phorbol myristate acetate (PMA), and the cultures were inoculated with E. faecalis for up to 48 h. At three time-points 90 min, 24 and 48 h after inoculation, the macrophages and their supernatants were examined. Assays included macrophage phagocytosis rate and vitality, bacterial survival, reactive oxygen species (ROS) production, mitochondrial activity, cytokine production and the expression of pro/anti-inflammatory M1/M2 markers. Also, periapical tissue from apicectomy samples of human endodontically treated teeth were collected for histological and immunofluorescent analysis. Statistical differences were analysed with RM ANOVA. RESULTS: E. faecalis were phagocytized, and subsequently, most of the macrophages underwent apoptosis and necrosis. The small population of macrophages that remained vital after 48 h post-inoculation harboured surviving bacteria. Despite a reduction in the number of macrophages over time, the mitochondrial activity of the surviving macrophages remained constant and external ROS decreased, whereas internal ROS increased. During the infection, a shift to a M2 macrophage population at 48 h post-infection was observed; the results were similar to those obtained in periapical human tissue biopsies (p < .05). CONCLUSIONS: The study portrays a continuous non-resolved infection with E. faecalis and activation of macrophages that are polarized to the M2 pro-resolution phenotype.

Salivary inflammatory cytokines echo the low inflammatory burden in liver-transplanted children.

OBJECTIVES: The aims of this study were to compare the salivary cytokine profile, as a potential replacement for blood tests, in liver-transplanted children to that of a control group of healthy children, and to correlate the values of commonly tested laboratory blood tests to those of published blood values. METHODS: Liver-transplanted children, and a control group of healthy children of the same sex and age distribution, were recruited for the study. Saliva was collected at the same appointment for routine blood tests for the liver-transplanted children. Saliva was also collected from a control group of healthy children with similar age and sex distributions. Normal healthy blood values were extracted from the literature, for comparison. Cytokine levels in the saliva were quantified with ELISA. The analysis compared serum and saliva values between liver-transplanted and healthy children. In the serum, the values of albumin, GIT, GPT, GGT, CRP, WBC, neutrophils, and lymphocytes were examined, while the levels of IL-6, CXCL1, IL-1b, and IL-10 were measured in the saliva. RESULTS: Thirty liver-transplanted children and 30 healthy children were included in the study. Compared with published data for healthy children, the liver-transplanted group showed similar hepatic serum levels, yet reduced levels of serum inflammatory markers. Compared with the control group, in the transplanted group, the mean value of IL-6 was lower and the mean value of CXCL1 was similar. Interestingly, the anti-inflammatory IL-10 cytokine was lower in the transplanted group, while the pro-inflammatory IL-1ß cytokine was higher. CONCLUSION: The salivary inflammatory markers examined showed a similar pattern to the serum inflammatory values, though different markers were examined in the serum and saliva. CLINICAL RELEVANCE: The current study stresses the potential of oral fluids as an accessible biofluid, for use as a diagnostic substrate for systemic and oral diseases. TRIAL REGISTRATION: 0136-16-RMC, Registered on 01 March 2018.

Diabetes as a risk factor for periodontal disease-plausible mechanisms.

The present narrative review examines the scientific evidence of the biological mechanisms that may link periodontitis and diabetes, as a source of comorbidity. Publications regarding periodontitis and diabetes, in human, animals, and in vitro were screened for their relevance. Periodontal microbiome studies indicate a possible association between altered glucose metabolism in prediabetes and diabetes and changes in the periodontal microbiome. Coinciding with this, hyperglycemia enhances expression of pathogen receptors, which enhance host response to the dysbiotic microbiome. Hyperglycemia also promotes pro-inflammatory response independently or via the advanced glycation end product/receptor for advanced glycation end product pathway. These processes excite cellular tissue destruction functions, which further enhance pro-inflammatory cytokines expression and alteration in the RANKL/osteoprotegerin ratio, promoting formation and activation of osteoclasts. The evidence supports the role of several pathogenic mechanisms in the path of true causal comorbidity between poorly controlled diabetes and periodontitis. However, further research is needed to better understand these mechanisms and to explore other mechanisms.

Follicular eruption with folliculotropic lymphocytic infiltrates associated with anti-tumor necrosis factor alpha therapy: Report and study of 3 cases.

BACKGROUND: We have encountered three cases of follicular eruptions with folliculotropic infiltrates of non-atypical lymphocytes associated with anti-tumor necrosis factor alpha (TNF-α) therapy. METHODS: Three patients aged 15 to 56 years treated with anti-TNF-α therapy (one with adalimumab, and two with infliximab) developed follicular eruptions characterized histopathologically by folliculotropic lymphocytic infiltrates. These were studied clinically, histopathologically, immunophenotypically, and molecularly. RESULTS: All three cases were characterized histopathologically by folliculotropic cell infiltrates of non-atypical T (CD3+) lymphocytes with variable follicular exocytosis. Marked reduction in CD7 staining and marked predominance of CD4+ cells over CD8+ cells were observed in 1 and 2 cases, respectively. T-cell receptor (TCR) gene rearrangement studies were monoclonal in 1 case. Discontinuation of anti-TNF-α therapy in all three cases, with corticosteroid creams in 1 case, led to complete resolution. Rechallenge with adalimumab in 1 case resulted in exacerbation. Replacement of therapy with non-anti-TNF-α biologic agents in 2 cases was not associated with recurrence. CONCLUSION: Follicular eruptions with folliculotropic lymphocytic infiltrates associated with anti-TNF-α therapy may show some immunophenotypical variations and/or monoclonal TCR gene rearrangements but lack sufficient cytomorphological features of folliculotropic MF. They may resolve with discontinuation of anti-TNF-α therapy.

The efficacy of pocket elimination/reduction compared to access flap surgery: A systematic review and meta-analysis.

AIM: To assess the efficacy and adverse effects of resective surgery compared to access flap in patients with periodontitis. METHODS: Randomized controlled trials with a follow-up ≥6 months were identified in ten databases. Screening, data extraction, and quality assessment were conducted by two reviewers. The primary outcome was probing pocket depth, and the main secondary outcome was clinical attachment level. Data on adverse events were collected. Meta-analysis was used to synthesize the findings of trials. RESULTS: A total of 880 publications were identified. Fourteen publications from nine clinical trials met the inclusion criteria and were included for analysis. Meta-analysis was carried out using all available results. The results indicated superior pocket depth reduction following resective surgery compared to access flap after 6-12 months of follow-up (weighted mean difference 0.47 mm; confidence interval 0.7-0.24; p = .010). After 36-60 months of follow-up, no differences were found between the two treatments in pocket depth and attachment level. The prevalence of adverse effects was not different between the groups. Post-operative recession tended to be more severe for the resective approaches. CONCLUSION: Resective surgical approach was superior to access flap in reducing pocket depth 6-12 months post-surgery, while no differences between the two modalities were found at 36-60 months of follow-up.

Follicular Eruption With Folliculotropic Lymphocytic Infiltrates Associated With Iatrogenic Immunosuppression: Report and Study of 3 Cases, and Review of the Literature.

BACKGROUND: Several cases of folliculotropic mycosis fungoides, associated with immunosuppressive therapy, including calcineurin inhibitors, have been reported in solid organ transplant patients. We have encountered 3 patients on immunosuppressive therapy who developed follicular eruptions with folliculocentric infiltrates of nonatypical lymphocytes. OBJECTIVE: To characterize these follicular eruptions and review the literature. METHODS: Three patients, aged 7-15 years, who were treated with systemic immunosuppressive therapy developed follicular eruptions characterized histopathologically by folliculocentric lymphocytic infiltrates. These were studied clinically, histopathologically, immunophenotypically, and molecularly for T-cell receptor (TCR) gene rearrangement. RESULTS: All 3 cases were characterized histopathologically by folliculocentric infiltrates of nonatypical CD3 T lymphocytes with variable follicular exocytosis. Two cases also showed follicular mucinosis. Marked reduction in CD7 staining, and marked predominance of CD4 cells over CD8 cells was observed in all 3 cases. The TCR gene rearrangement studies were monoclonal in 2 cases. Oral calcineurin inhibitors (2 cyclosporine A and 1 tacrolimus) were part of the therapeutic regimen in all 3 patients. Their cessation along with local corticosteroid creams in 2 patients, and phototherapy with oral acitretin in one patient, was associated with complete clinical remission. CONCLUSIONS: Patients undergoing systemic immunosuppressive therapy that includes calcineurin inhibitors might develop follicular eruption with some immunophenotypical variations and a monoclonal TCR gene rearrangement but lack sufficient cytomorphological features of folliculotropic mycosis fungoides. Altering the immunosuppressive agent including calcineurin inhibitors may result in regression of the eruptions.

[DERMATOLOGIC CONDITIONS IN MONOZYGOTIC TWINS].

INTRODUCTION: Genetic twin studies may shed light on the genetic basis as well as environmental and epigenetic factors in disease pathogenesis. Herein, we present four pairs of monozygotic twins sharing similar phenotypes in three dermatologic conditions, and a literature review regarding twin studies in these diseases.

High-resolution novel method for tracking bacteria in a multi-species biofilm.

The aim of this study is to establish a novel high resolution tracking ability of a specific bacterium in multispecies biofilm. A periodontal multispecies biofilm was constructed with Streptococcus sanguis, Actinomyces naeslundii, Porphyromonas gingivalis and Fusobacterium nucleatum. A single species was stained with fluorescein isothiocyanate (FITC). The mature biofilm was stained for viability (propidium iodide) and analysis was performed with flow cytometry. The sensitivity of the assay was compared with colony forming units (CFU) counts. A single cell suspension of P. gingivalis was grown in broth and biofilm to identify the location of these events on side scatter and forward scatter. The sensitivity of the assay was comparable to that of the CFU counts. The assay allows quantification of the ratio of a single bacterium within the biofilm, and its viable proportion. The described method is reproducible and of high resolution, and allows the examination of microbes' composition and viability within a biofilm structure.

Myd88 plays a major role in the keratinocyte response to infection with Porphyromonas gingivalis.

AIM: To explore the role of keratinocyte myeloid differentiation primary response 88 (MyD88) expression in the adhesion of Porphyromonas gingivalis to the cells and its subsequent invasion and intracellular survival. MATERIALS AND METHODS: Primary mouse keratinocytes from wild-type (WT) or Myd88-/- mice were infected with P gingivalis alone or co-infected with Fusobacterium nucleatum. Bacterial adhesion and invasion were measured using fluorescent microscopy and flow cytometry, and intracellular survival in keratinocytes was quantified by an antibiotic protection assay. Keratinocyte expression of antimicrobial peptides was measured by real-time PCR. RESULTS: In the absence of MyD88, P gingivalis adherence, invasion, and intracellular survival were enhanced compared with WT keratinocytes. The presence of F nucleatum during infection increased the adhesion of P gingivalis to WT keratinocytes but reduced the adhesion to Myd88-/- keratinocytes. Fusobacterium nucleatum improved mildly the invasion and survival of P gingivalis in both cell types. Baseline expression of beta-defensin 2, 3, 4 and RegIII-γ was elevated in Myd88-/- keratinocytes compared to WT cells; however, following infection beta-defensin expression was strongly induced in WT cells but decreased dramatically in the MyD88 deficient cells. CONCLUSION: In the absence of MyD88 expression, P gingivalis adhesion to keratinocytes is improved, and invasion and intracellular survival are increased. Furthermore, keratinocyte infection by P gingivalis induces antimicrobial peptide expression in a MyD88-dependent manner. Thus, MyD88 plays a key role in the interaction between P gingivalis and keratinocytes.

Incorporating antibiotics into platelet-rich fibrin: A novel antibiotics slow-release biological device.

BACKGROUND: The aim of the present in vitro study was to explore the possibility of using platelet-rich fibrin (PRF) as a local sustained released device for antibiotics. MATERIALS AND METHODS: Platelet-rich fibrin was prepared with the addition of antibiotics (5 mg/ml metronidazole; 150 mg/ml clindamycin; 1 mU/ml penicillin) or saline prior to centrifugation, while collagen sponges served as control. PRFs anti-bacterial properties were examined in an anti-biogram assay with Staphylococcus aureus or Fusobacterium nucleatum at different time intervals after PRF preparation. RESULTS: The addition of antibiotic solutions at volumes of 2 or 1 ml led to significant changes in PRF's physical properties, while the addition of 0.5 ml solution did not. PRF with saline showed minor anti-bacterial activity, while all PRFs with antibiotics showed significant anti-bacterial activity (p < 0.05). No differences were observed between raw (clot) and pressed (membrane) forms of PRF. Collagen sponges with and without antibiotics showed similar results to PRF. PRF and collagen sponges with antibiotics preserved their anti-bacterial properties 4 days after preparation. CONCLUSIONS: Platelet-rich fibrin incorporated with antibiotics showed long-term anti-bacterial effect against F. nucleatum and S. aureus. This modified PRF preparation may be used to reduce the risk of post-operative infection in addition to the beneficial healing properties of PRF.

Nupharidine enhances Aggregatibacter actinomycetemcomitans clearance by priming neutrophils and augmenting their effector functions.

OBJECTIVES: Nupharidine (6,6'-Dihydroxythiobinupharidine), purified from the aquatic plant Nuphar lutea leaves (Water lily) prompts antimicrobial activity of immune cells. The aim of the study was to test the effect of Nupharidine on neutrophil function against Aggregatibacter actinomycetemcomitans, JP2 clone (Aa-JP2). METHODS: Neutrophils derived from the human cell line HL60 and human peripheral blood derived from aggressive periodontitis and periodontally healthy subjects were incubated with Nupharidine or vehicle and inoculated with JP2. Bacterial survival was tested using viable counts on blood agar (CFU's). Neutrophils' necrosis/apoptosis, reactive oxygen species (ROS) production, phagocytosis and neutrophil extracellular traps (NET) production following infection were tested, as well as markers of neutrophil priming. RESULTS: Nupharidine had no direct bactericidal effect on JP2, but it enhanced Aa-JP2 clearance by neutrophils. Nupharidine enhanced neutrophil phagocytosis, ROS production and NET formation during JP2 infection. Furthermore, Nupharidine enhanced the expression of certain markers of neutrophils priming, specifically iCAM1, DECTIN-2 and intracellular IL-1ß. CONCLUSION: Nupharidine was shown to promote neutrophil effector bactericidal functions, boosting Aa-JP2 clearance. The results point to the potential of Nupharidine as an adjunctive agent in the treatment of Aa-JP2 periodontitis, but this should be tested initially using pre-clinical and clinical studies.

Indirect Bactericidal Properties of Recombinant Human Bone Morphogenetic Protein 2 In Vitro.

PURPOSE: Bone morphogenetic proteins (BMPs) are secreted cytokines and are involved in various metabolic functions and inflammatory processes in different organs. The purpose of this study was to investigate whether BMPs also possess antimicrobial properties in direct or indirect ways. MATERIALS AND METHODS: Antibacterial properties of recombinant human BMP2 (rhBMP2) were tested on 4 bacteria species (Staphylococcus aureus, Escherichia coli, Streptococcus mitis, Streptococcus constellatus) to examine the potential synergism of rhBMP2 with antibiotics. Indirect antibacterial properties were tested by infecting neutrophils with rhBMP2 and bacteria to investigate bacterial survival. Reactive oxidative species (ROS) production in neutrophils in the presence of rhBMP2 also was tested. RESULTS: RhBMP2 in cardboard disks or sponge collagen as carriers did not show antibacterial activity against all tested bacteria. Further, synergism of rhBMP2 with antibiotics was not evident. Survival of bacteria inoculated with neutrophils and rhBMP2 led to a marked decrease in bacterial survival compared with neutrophils without rhBMP2. Although rhBMP2 inoculation of neutrophils alone did not induce ROS, its presence with the bacterial infection showed augmented ROS production for all tested bacteria. CONCLUSIONS: RhBMP2 did not show direct antibacterial properties but did exhibit an indirect bactericidal effect in the presence of neutrophils. ROS production indicated that rhBMP2 has a role as a priming agent for neutrophils by augmenting their bactericidal capabilities and suggests the importance of its presence in contaminated surgical bone augmentation sites.

Development of In-Browser Simulators for Medical Education: Introduction of a Novel Software Toolchain.

BACKGROUND: Simulators used in teaching are interactive applications comprising a mathematical model of the system under study and a graphical user interface (GUI) that allows the user to control the model inputs and visualize the model results in an intuitive and educational way. Well-designed simulators promote active learning, enhance problem-solving skills, and encourage collaboration and small group discussion. However, creating simulators for teaching purposes is a challenging process that requires many contributors including educators, modelers, graphic designers, and programmers. The availability of a toolchain of user-friendly software tools for building simulators can facilitate this complex task. OBJECTIVE: This paper aimed to describe an open-source software toolchain termed Bodylight.js that facilitates the creation of browser-based client-side simulators for teaching purposes, which are platform independent, do not require any installation, and can work offline. The toolchain interconnects state-of-the-art modeling tools with current Web technologies and is designed to be resilient to future changes in the software ecosystem. METHODS: We used several open-source Web technologies, namely, WebAssembly and JavaScript, combined with the power of the Modelica modeling language and deployed them on the internet with interactive animations built using Adobe Animate. RESULTS: Models are implemented in the Modelica language using either OpenModelica or Dassault Systèmes Dymola and exported to a standardized Functional Mock-up Unit (FMU) to ensure future compatibility. The C code from the FMU is further compiled to WebAssembly using Emscripten. Industry-standard Adobe Animate is used to create interactive animations. A new tool called Bodylight.js Composer was developed for the toolchain that enables one to create the final simulator by composing the GUI using animations, plots, and control elements in a drag-and-drop style and binding them to the model variables. The resulting simulators are stand-alone HyperText Markup Language files including JavaScript and WebAssembly. Several simulators for physiology education were created using the Bodylight.js toolchain and have been received with general acclaim by teachers and students alike, thus validating our approach. The Nephron, Circulation, and Pressure-Volume Loop simulators are presented in this paper. Bodylight.js is licensed under General Public License 3.0 and is free for anyone to use. CONCLUSIONS: Bodylight.js enables us to effectively develop teaching simulators. Armed with this technology, we intend to focus on the development of new simulators and interactive textbooks for medical education. Bodylight.js usage is not limited to developing simulators for medical education and can facilitate the development of simulators for teaching complex topics in a variety of different fields.

Microbial accumulation on different suture materials following oral surgery: a randomized controlled study.

BACKGROUND: The aim of the study was to compare bacterial accumulation on different suture materials following oral surgery. METHODS: Patients scheduled for implant or periodontal surgery were included in the study. Upon flap closure, four different sutures were placed in a randomized sequence-silk, coated polyglactin, nylon, and polyester. Ten days following surgery, the sutures were removed and incubated in aerobic as well as anaerobic conditions for 7 days and colony-forming units (CFUs) were calculated. Association between bacterial accumulation and periodontal diagnosis, type of surgery, and antibiotic treatment were also tested. RESULTS: All sutures in all patients were found to contain bacteria. Overall, nylon sutures showed significantly lower CFU levels compared to silk, coated polyglactin, and polyester sutures. The type of surgery (implant vs. periodontal surgery) did not significantly influence bacterial accumulation. Also, periodontal diagnosis had little impact on CFU counts. Interestingly, post-surgical antibiotic treatment also had only a minor effect on bacterial accumulation on the various sutures. DISCUSSION: The results indicate that the monofilamentous nylon sutures showed less microbial accumulation than the other tested materials that were all braided. This effect may be due to material qualities as well as suture macrostructure. Type of surgery, periodontal diagnosis, and antibiotic consumption have little effect on bacterial accumulation of sutures. CLINICAL RELEVANCE: The study provides the microbial profile of commonly used sutures and may assist in suture selection during clinical procedures.

An update on the evidence for pathogenic mechanisms that may link periodontitis and diabetes.

AIM: To provide an update of the review by Taylor (Journal of Clinical Periodontology, 2013, 40, S113) regarding the scientific evidence of the biological association between periodontitis and diabetes. METHODS: Literature searches were performed using MeSH terms, keywords and title words and were published between 2012 and November 2016. All publications were screened for their relevance. The data from the articles were extracted and summarized in tables and a narrative review. RESULTS: Small-scale molecular periodontal microbiome studies indicate a possible association between altered glucose metabolism in pre-diabetes and diabetes and changes in the periodontal microbiome, with no evidence for casual relationships. Clinical and animal studies found elevated gingival levels of IL1-ß, TNF-α, IL-6, RANKL/OPG and oxygen metabolites in poorly controlled diabetes. In addition, individuals with diabetes and periodontitis exhibit high levels of circulating TNF-α, CRP and mediators of oxidative stress, and successful periodontal treatment reduces their levels. CONCLUSIONS: The elevated pro-inflammatory factors in the gingiva of patients with poorly controlled diabetes suggest a biological pathway that may aggravate periodontitis. Some evidence suggests that the systemic inflammatory burden in periodontitis has the potential to affect diabetes control, but no studies addressed the impact of successful periodontal therapy on the pathophysiological mechanisms involved in systemic complications of diabetes.

The Role of Histopathology and Immunohistochemistry in the Diagnosis of Cutaneous Leishmaniasis Without "Discernible" Leishman-Donovan Bodies.

BACKGROUND: Histopathology plays an important role in the diagnosis of cutaneous leishmaniasis (CL) but Leishman-Donovan (LD) bodies may not always be discernible. Recently, anti-CD1a antibody (Ab), clone MTB1, was found to decorate LD bodies immunohistochemically. OBJECTIVE: Can histopathology without discernible LD bodies be used to diagnose CL, and can immunohistochemistry using anti-CD1a Ab, clone MTB1, detect LD bodies in these cases. METHODS: Suspected CL lesions were studied histopathologically and immunohistochemically, and the patients' clinical files were reviewed. RESULTS: Of the 196 patients with suspected CL, direct smear demonstrated LD bodies in 50 (25.5%). Of the remaining 146 patients, 118 underwent biopsy. In 56 (47.5%) patients, the hematoxylin-eosin-stained sections revealed LD bodies. In 47 (39.8%) patients, LD bodies were not discerned but the histopathology demonstrated histiocytic infiltrates with varying numbers of plasma cells along with other inflammatory cells, and negative Ziehl-Neelsen and periodic acid-Schiff stains. This pattern was termed "histopathology consistent with leishmaniasis." The history, clinical findings, and response to anti-leishmania therapy supported the diagnosis of CL in all of them, and immunostains for CD1a, clone MTB1, detected LD bodies in 11 (23.4%) of these 47 patients. CONCLUSIONS: "Histopathology consistent with CL" along with appropriate clinical findings supports the diagnosis of CL in an endemic area, and immunostains with CD1a Ab, clone MTB1, may help in the minority of the cases.

Antibacterial Orthodontic Adhesive Incorporating Polyethyleneimine Nanoparticles.

PURPOSE: To evaluate the antibacterial, mechanical and biocompatibility characteristics of an orthodontic adhesive that contains quaternary ammonium polyethyleneimine (QPEI) nanoparticles. MATERIALS AND METHODS: QPEI nanoparticles were added to an orthodontic adhesive at 0%, 1% and 1.5% wt/wt. Antibacterial activity was tested after aging for 14 days using the direct contact test (DCT). The degree of monomer conversion (DC) was measured using Fourier transform infrared (FTIR) spectroscopy. Shear bond atrength (SBS) was tested on the etched enamel of extracted human teeth. Biocompatibility was tested using keratinocyte and neutrophil cell lines in the XTT assay. RESULTS: The DCT results showed significant bacterial growth inhibition in the test group incorporating 1.5% wt/wt QPEI nanoparticles (p < 0.05). The DC of the 0%, 1%, and 1.5% wt/wt samples measured immediately and after 10 min was 62.2-71.0%, 59.1-68.7%, and 52.9-58.6%, respectively, and the average SBS were 9.25 MPa, 11.57 MPa, and 9.10 MPa, respectively. Keratinocyte and neutrophil viability did not change following the addition of QPEI to the orthodontic adhesives. CONCLUSIONS: The incorporation of QPEI nanoparticles into orthodontic cement provides long-lasting antibacterial activity against Streptococcus mutans without reducing the strength of adhesion to enamel, the degree of double bond conversion during the polymerisation, or the biocompatibility of the orthodontic cement.

Are anti-inflammatory agents effective in treating gingivitis as solo or adjunct therapies? A systematic review.

OBJECTIVE: Systematically review the scientific evidence for efficiency of anti-inflammatory agents against gingivitis, either as solo treatments or adjunctive therapies. METHODS: A protocol was developed aimed to answer the following focused question: "Are anti-inflammatory agents effective in treating gingivitis as solo or adjunct therapies?" RCTs and cohort studies on anti-inflammatory agents against gingivitis studies were searched electronically. Screening, data extraction and quality assessment were conducted. The primary outcome measures were indices of gingival inflammation. A sub-analysis was performed dividing the active agents into anti-inflammatory and other drugs. RESULTS: The search identified 3188 studies, of which 14 RCTs met the inclusion criteria. The use of anti-inflammatory or other agents, in general showed a higher reduction in the test than in the control in terms of gingival indexes and bleeding scores. Only two RCTs on inflammatory drugs could be meta-analysed, showing a statistically significant reduction in the GI in the experimental group [WMD = -0.090; 95% CI (-0.105; -0.074); p = 0.000]. However, the contribution of both studies to the global result was unbalanced (% weight: 99.88 and 0.12 respectively). CONCLUSIONS: Most of the tested material showed beneficial effect as anti-inflammatory agents against gingivitis, either as a single treatment modality or as an adjunctive therapy.